Effects of oral probiotics administration on the expression of transforming growth factor ß and the proinflammatory cytokines interleukin 6, interleukin 17, and tumor necrosis factor α in skin wounds in rats.

BACKGROUND: Cytokines and growth factors play key roles during the tissue repair process. We aim to evaluate the effect of perioperative oral of probiotics, on the healing process in skin wound in rats, by histological aspects, and by the expression of TGF-ß, and the pro-inflammatory cytokines IL6, IL7, and TNF-α. METHODS: 72 adult male Wistar rats were split into two groups control (n = 36) and probiotic group (n = 36). Each group was subdivided into three subgroups with 12 animals each according to euthanasia day: 3rd, 7th, and 10th postoperative(PO) day. RESULTS: Wound contraction was faster with the use of probiotics (p = .013). Also fibrosis was significantly higher in the Probiotic group in the 7th PO day (p = .028). In the probiotic group, there was a reduction of TNF-α at 3th PO day (p = .023); and a reduction of IL6 in 7th PO day (p = .030). There was also a reduction of the expression of IL-17 in 3rd PO day (p = .039) and 7rd PO day (P = .024). In contrast, TGF-ß was lower in the 10th PO day (p = .031) in the probiotic group as compared to controls, indicating that the increase of the fibrosis caused negative feedback with the TGF-ß. CONCLUSION: Probiotics are associated with a shorter inflammatory phase by attenuating the expression of cytokines IL-6 and TNF-α and accelerating the reduction of IL-17 and TGF-ß, leading to faster and improved cutaneous healing in rats.

EFFECTS OF PROBIOTICS SUPPLEMENTATION ON SKIN WOUND HEALING IN DIABETIC RATS.

BACKGROUND: Example of wound contraction area at: A) day of surgery in the control group; B) 7PO in the control group; C) day of surgery in the probiotic group; D) 7PO in probiotic group. Chronic wounds in patients with Diabetes Mellitus often become incurable due to prolonged and excessive production of inflammatory cytokines. The use of probiotics modifies the intestinal microbiota and modulates inflammatory reactions. AIM: To evaluate the influence of perioperative supplementation with probiotics in the cutaneous healing process in diabetic rats. METHODS: Forty-six rats were divided into four groups (C3, P3, C10, P10) according to the treatment (P=probiotic or C=control, both orally administered) and day of euthanasia, 3rd or 10th postoperative days. All rats were induced to Diabetes Mellitus 72 h before starting the experiment with alloxan. Supplementation was initiated five days before the incision and maintained until euthanasia. Scalpel incision was guided by a 2x2 cm mold and the wounds were left to heal per second-intention. The wounds were digitally measured. Collagen densitometry was done with Picrosirius Red staining. Histological parameters were analyzed by staining by H&E. RESULTS: The contraction of the wound was faster in the P10 group which resulted in a smaller scar area (p=0.011). There was an increase in type I collagen deposition from the 3rd to the 10th postoperative day in the probiotic groups (p=0.016), which did not occur in the control group (p=0.487). The histological analysis showed a better degree of healing in the P10 group (p=0.005), with fewer polymorphonuclear (p<0.001) and more neovessels (p=0.001). CONCLUSIONS: Perioperative supplementation of probiotics stimulates skin wound healing in diabetic rats, possibly due to attenuation of the inflammatory response and increased neovascularization and type I collagen deposition.

EFFECT OF PROBIOTIC ORAL ADMINISTRATION ON SKIN WOUND HEALING IN RATS.

BACKGROUND: Manipulating intestinal microbiota with probiotics might stimulate skin response. Understanding all stages of the healing process, as well as the gut-skin-healing response can improve the skin healing process. AIM: To evaluate the effect of perioperative oral administration of probiotics on the healing of skin wounds in rats. METHODS: Seventy-two Wistar male adult rats were weighed and divided into two groups with 36 each, one control group (supplemented with oral maltodextrin 250 mg/day) and one probiotic group (supplemented with Lactobacillus paracasei LPC-37, Bifidobacterium lactis HN0019, Lactobacillus rhamnosus HN001, Lactobacillus acidophilus NCFM® at a dose of 250 mg/day), both given orally daily for 15 days. The two groups were subsequently divided into three subgroups according to the moment of euthanasia: in the 3rd, 7th and 10th postoperative days. RESULTS: There were no significant changes in weight in both groups. Wound contraction was faster in probiotic group when compared to the controls, resulting in smaller wound area in the 7th postoperative day. As for histological aspects, the overall H&E score was lower in the probiotic group. The probiotic group showed increased fibrosis from 3rd to the 7th postoperative day. The type I collagen production was higher in the probiotic group at the 10th postoperative day, and the type III collagen increased in the 7th. CONCLUSION: The perioperative use of orally administrated probiotic was associated with a faster reduction of the wound area in rats probably by reducing the inflammatory phase, accelerating the fibrosis process and the deposition of collagen.

THE IMPACT OF THE USE OF SYMBIOTICS IN THE PROGRESSION OF NONALCOHOLIC FATTY LIVER DISEASE IN A RAT MODEL.

BACKGROUND: Non-alcoholic fatty liver disease (NAFLD) is characterized by accumulation of intrahepatic lipid. The use of live microorganisms promotes beneficial effects; however, the use of symbiotic and its role in NAFLD is not yet fully understood. AIM: Verify if the symbiotic administration influences the occurrence and progression of NAFLD in rats, after induction of hepatic steatosis by high calorie diet. METHOD: Forty-five rats were divided into four groups: G1 (control); G2 (control+symbiotic); G3 (high calorie+symbiotic) and G4 (high calorie), and euthanized after 60 days of diet. Liver disease was evaluated by biochemical analysis, IL6 measurement and histological assessment. RESULTS: Symbiotic had influence neither on weight gain, nor on coefficient dietary intake in G3 and G4. G2 had the greatest weight gain, while G1 had the highest coefficient dietary intake between groups. G1 showed higher expression of aspartate aminotransferase than those from G2 (150±35 mg/dl, and 75±5 mg/dl) while G4 showed higher expression of the enzyme compared to G3 (141±9.7 mg/dl to 78±4 mg/dl). Liver histology showed different stages of NAFLD between groups. G4 animals showed increased serum interleukin-6 when compared to G3 (240.58±53.68 mg/dl and 104.0±15.31 mg/dl). CONCLUSION: Symbiotic can reduce hepatic aminotransferases and interleukin-6 expression. However, the histology showed that the symbiotic was not able to prevent the severity of NAFLD in rats.

Effects of probiotics supplementation on skin wound healing in diabetic rats / Suplementação perioperatória com probióticos na cicatrização de feridas cutâneas em ratos diabéticos

ABSTRACT Background: Chronic wounds in patients with Diabetes Mellitus often become incurable due to prolonged and excessive production of inflammatory cytokines. The use of probiotics modifies the intestinal microbiota and modulates inflammatory reactions. Aim: To evaluate the influence of perioperative supplementation with probiotics in the cutaneous healing process in diabetic rats. Methods: Forty-six rats were divided into four groups (C3, P3, C10, P10) according to the treatment (P=probiotic or C=control, both orally administered) and day of euthanasia, 3rd or 10th postoperative days. All rats were induced to Diabetes Mellitus 72 h before starting the experiment with alloxan. Supplementation was initiated five days before the incision and maintained until euthanasia. Scalpel incision was guided by a 2x2 cm mold and the wounds were left to heal per second-intention. The wounds were digitally measured. Collagen densitometry was done with Picrosirius Red staining. Histological parameters were analyzed by staining by H&E. Results: The contraction of the wound was faster in the P10 group which resulted in a smaller scar area (p=0.011). There was an increase in type I collagen deposition from the 3rd to the 10th postoperative day in the probiotic groups (p=0.016), which did not occur in the control group (p=0.487). The histological analysis showed a better degree of healing in the P10 group (p=0.005), with fewer polymorphonuclear (p<0.001) and more neovessels (p=0.001). Conclusions: Perioperative supplementation of probiotics stimulates skin wound healing in diabetic rats, possibly due to attenuation of the inflammatory response and increased neovascularization and type I collagen deposition.

RESUMO Racional: Feridas crônicas em pacientes diabéticos muitas vezes se tornam incuráveis devido à produção prolongada e excessiva de citocinas inflamatórias. A utilização de probióticos modifica a microbiota intestinal e modula reações inflamatórias. Objetivo: Avaliar a influência da suplementação perioperatória com probióticos no processo de cicatrização cutânea em ratos diabéticos. Método: Quarenta e seis ratos foram divididos em quatro grupos (C3, P3, C10, P10) conforme tratamento (P=probiótico ou C=controle, via oral) e dia de eutanásia: 3o ou 10o dia de pós-operatório. Todos os ratos foram induzidos ao diabete melito 72 h antes de iniciar o experimento com aloxana. A suplementação foi iniciada cinco dias antes da operação e mantida até a eutanásia. Foi realizada incisão com bisturi guiada por molde de 2x2 cm e a ferida foi deixada para cicatrizar por segunda intenção. As feridas foram medidas digitalmente. A densitometria de colágeno foi determinada com coloração picrosirius red. A histologia foi avaliada por coloração com H&E. Resultados: A contração da ferida foi maior no grupo P10, o que resultou em menor área cruenta (p=0,011). Houve aumento do colágeno tipo I do 3o para o 10o dia de pós-operatório no grupo P10 (p=0,016), o que não ocorreu no grupo controle (p=0,487). A análise histológica mostrou melhor grau de cicatrização no grupo P10 (p=0,005), com menos polimorfonucleares (p<0,001) e mais neovasos (p=0,001). Conclusões: A suplementação perioperatória de probióticos promove aceleração da cicatrização cutânea em ratos diabéticos, possivelmente por atenuar a resposta inflamatória e aumentar a neovascularização e a deposição de colágeno tipo I.

The impact of the use of symbiotics in the progression of nonalcoholic fatty liver disease in a rat model / Impacto do uso de simbiótico na doença hepática gordurosa não alcoólica em modelo de ratos

ABSTRACT Background: Non-alcoholic fatty liver disease (NAFLD) is characterized by accumulation of intrahepatic lipid. The use of live microorganisms promotes beneficial effects; however, the use of symbiotic and its role in NAFLD is not yet fully understood. Aim: Verify if the symbiotic administration influences the occurrence and progression of NAFLD in rats, after induction of hepatic steatosis by high calorie diet. Method: Forty-five rats were divided into four groups: G1 (control); G2 (control+symbiotic); G3 (high calorie+symbiotic) and G4 (high calorie), and euthanized after 60 days of diet. Liver disease was evaluated by biochemical analysis, IL6 measurement and histological assessment. Results: Symbiotic had influence neither on weight gain, nor on coefficient dietary intake in G3 and G4. G2 had the greatest weight gain, while G1 had the highest coefficient dietary intake between groups. G1 showed higher expression of aspartate aminotransferase than those from G2 (150±35 mg/dl, and 75±5 mg/dl) while G4 showed higher expression of the enzyme compared to G3 (141±9.7 mg/dl to 78±4 mg/dl). Liver histology showed different stages of NAFLD between groups. G4 animals showed increased serum interleukin-6 when compared to G3 (240.58±53.68 mg/dl and 104.0±15.31 mg/dl). Conclusion: Symbiotic can reduce hepatic aminotransferases and interleukin-6 expression. However, the histology showed that the symbiotic was not able to prevent the severity of NAFLD in rats.

RESUMO Racional: A doença hepática gordurosa não alcoólica (DHGNA) é caracterizada por acúmulo de lipídios intra-hepáticos. O uso de microrganismos vivos promove diversos efeitos benéficos; porém, a utilização de simbióticos e sua atuação na DHGNA ainda não está totalmente esclarecida. Objetivo: Verificar se a administração de simbióticos influencia na ocorrência e na progressão da DHGNA em ratos, após a indução de esteatose hepática por dieta hipercalórica. Método: Quarenta e cinco ratos Wistar foram divididos em quatro grupos: G1 (controle); G2 (controle+simbiótico); G3 (hipercalórica+ simbiótico) e G4 (hipercalórica), e eutanasiados após 60 dias de dieta. Coleta de sangue foi realizada para obtenção de análises bioquímicas e dosagem de IL6, e de tecido para análise histológica do fígado. Resultados: O simbiótico não influenciou no ganho de peso e no coeficiente de consumo alimentar nos grupos G3 e G4. Já G2 obteve maior ganho de peso, enquanto G1 apresentou o maior coeficiente de consumo alimentar entre os grupos. G1 apresentou maior expressão de aspartato aminotransferase em relação ao G2 (150±35 mg/dl e 75±5 mg/dl), enquanto G4 teve maior expressão desta enzima em relação ao G3 (141±9,7 mg/dl e 78±4 mg/dl,). A análise histológica hepática mostrou diferentes estágios de evolução da DHGNA entre os grupos. Animais do G4 apresentaram aumento sérico de interleucina-6 quando comparados a G3 (240,58±53,68 mg/dl, e 104,0±15,31 mg/dl). Conclusão: Os simbióticos reduziram aminotransferases hepáticas e a expressão de interleucina-6. No entanto, a histopatologia demonstrou que o simbiótico não foi capaz de prevenir a severidade da DHGNA em ratos.